Molecular barcoding on opposite transcript ends

Abstract

Disclosed herein include systems, methods, compositions, and kits for molecular barcoding on the 5′-end of a nucleic acid target. After barcoding a nucleic acid target using an oligonucleotide barcode comprising a target binding region and a molecular label to generate a barcoded nucleic acid molecule, an oligonucleotide comprising a complement of the target binding region can be added to generate a barcoded nucleic acid molecule comprising the target-binding region and the complement of the target-binding region. A stem loop is formed with intra-molecular hybridization of the barcoded nucleic acid molecule, which can be extended to generate an extended barcoded nucleic acid molecule comprising the molecular label and a complement of the molecular label.

Description

RELATED APPLICATIONS[0001]The present application claims the benefit of U.S. Provisional Application No. 62 / 666,506, filed May 3, 2018, which is hereby incorporated by reference in its entirety.FIELD[0002]The present disclosure relates generally to the field of molecular biology, and for particular to multiomics analyses using molecular barcoding.Description of the Related Art[0003]Methods and techniques such as molecular barcoding are useful for single cell transcriptomics analysis, including deciphering gene expression profiles to determine the states of cells using, for example, reverse transcription, polymerase chain reaction (PCR) amplification, and next generation sequencing (NGS). Molecular barcoding are also useful for single cell proteomics analysis.SUMMARY[0004]Disclosed herein includes methods for attaching oligonucleotide barcodes to a target in a sample. By way of example, the target can comprise, consist essentially of, or consist of a nucleic acid target. In some embo...

AI Technical Summary

Benefits of technology

[0025]Disclosed herein includes kits for attaching oligonucleotide barcodes to a target in a sample, determining the numbers of targets in a sample, and/or determining the numbers of a nucleic acid target in a sample. In some embodiments, the kit includes: a plurality of oligonucleotide barcodes, wherein each of the plurality of oligonucleotide barcodes comprises a molecular label and a target-binding region, and wherein at least 10 of the plurality of oligonucleotide barcodes comprise different molecular label sequences; a terminal deoxynucleotidyl transferase or a ligase; and a DNA polymerase lacking at least one...

Problems solved by technology

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